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Literature summary for 2.1.1.151 extracted from

  • Frank, S.; Deery, E.; Brindley, A.A.; Leech, H.K.; Lawrence, A.; Heathcote, P.; Schubert, H.L.; Brocklehurst, K.; Rigby, S.E.; Warren, M.J.; Pickersgill, R.W.
    Elucidation of substrate specificity in the cobalamin (vitamin B12) biosynthetic methyltransferases. Structure and function of the C20 methyltransferase (CbiL) from Methanothermobacter thermautotrophicus (2007), J. Biol. Chem., 282, 23957-23969.
    View publication on PubMed
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Cloned(Commentary)

Cloned (Comment) Organism
complementation studies with the Salmonella enterica mutant strain AR3711, wild type but non of the mutant proteins complements enzyme deficiency of the host strain Methanothermobacter thermautotrophicus
wild type and mutant proteins expressed as His-tag fusion protein in Escherichia coli Rosetta(DE3)pLysS Methanothermobacter thermautotrophicus

Crystallization (Commentary)

Crystallization (Comment) Organism
crystallized by hanging drop vapor diffusion method in the presence of S-adenosyl-L-methionine or S-adenosyl-L-homocysteine, crystals complemented with substrate by soaking in substrate solution, no crystal growth in the absence of S-adenosyl-L-methionine or S-adenosyl-L-homocysteine Methanothermobacter thermautotrophicus

Protein Variants

Protein Variants Comment Organism
D104A inactive in vivo as shown by complementation studies with Salmonella enterica, low in vitro activity, reduced affinity for S-adenosyl-L-methionine Methanothermobacter thermautotrophicus
K176A inactive in vivo as shown by complementation studies with Salmonella enterica, no structural changes compared with crystallized wild-type enzyme Methanothermobacter thermautotrophicus
Y220A inactive in vivo as shown by complementation studies with Salmonella enterica Methanothermobacter thermautotrophicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29000
-
 2 * 29000, SDS-PAGE, recombinant His-tagged protein, native mass by gel filtration Methanothermobacter thermautotrophicus
51000
-
 gel filtration Methanothermobacter thermautotrophicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + cobalt-factor II Methanothermobacter thermautotrophicus key enzyme of the anaerobic cobalamin biosynthesis, important step in ring contraction S-adenosyl-L-homocysteine + cobalt-factor III
-
 ?

Organism

Organism UniProt Comment Textmining
Methanothermobacter thermautotrophicus O27402
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant proteins from Escherichia coli Methanothermobacter thermautotrophicus

Reaction

Reaction Comment Organism Reaction ID
S-adenosyl-L-methionine + cobalt-factor II = S-adenosyl-L-homocysteine + cobalt-factor III Gly11, Gly103 and Asp104 involved in S-adenosyl-L-methionine binding Methanothermobacter thermautotrophicus
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0041
-
 37°C Methanothermobacter thermautotrophicus
0.0136
-
 65°C Methanothermobacter thermautotrophicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information metal-free factor II does not serve as substrate Methanothermobacter thermautotrophicus ?
-
 ?
S-adenosyl-L-methionine + Co(II)-cobalt-factor II
-
 Methanothermobacter thermautotrophicus S-adenosyl-L-homocysteine + cobalt-factor III
-
 ?
S-adenosyl-L-methionine + Co(II)-precorrin-2 slow reaction with low product yield Methanothermobacter thermautotrophicus S-adenosyl-L-homocysteine + Co(II)-precorrin-3
-
 ?
S-adenosyl-L-methionine + cobalt-factor II
-
 Methanothermobacter thermautotrophicus S-adenosyl-L-homocysteine + cobalt-factor III
-
 ?
S-adenosyl-L-methionine + cobalt-factor II key enzyme of the anaerobic cobalamin biosynthesis, important step in ring contraction Methanothermobacter thermautotrophicus S-adenosyl-L-homocysteine + cobalt-factor III
-
 ?

Subunits

Subunits Comment Organism
dimer crystal structure analysis Methanothermobacter thermautotrophicus
dimer 2 * 29000, SDS-PAGE, recombinant His-tagged protein, native mass by gel filtration Methanothermobacter thermautotrophicus

Synonyms

Synonyms Comment Organism
CbiL belongs to the class III methyltransferases Methanothermobacter thermautotrophicus